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. Author manuscript; available in PMC: 2020 May 26.
Published in final edited form as: J Vis Exp. 2019 Oct 3;(152):10.3791/59977. doi: 10.3791/59977
Name of Material/Equipment Company Catalog Number Comments/Description
1/2 cc LO-DOSE INSULIN SYRINGE EXELINT 26028 general supply
293T cells ATCC CRL-3216™ Cell line
BD Luer-Lok disposable syringes, 10 ml BD 309604 general supply
BD Microtainer blood collection tubes, K2EDTA added BD Bioscience 365974 general supply
BD Precisionglide needle, 18 G BD 305195 general supply
BD Precisionglide needle, 22 G BD 305155 general supply
Collagen from calf skin Sigma-Aldrich 9007-34-5 general supply
Corning Costar Ultra-Low Attachment Multiple Well Plate, 6 well Millipore Sigma CLS3471 general supply
CRISPR/Cas9 knock-in mice The Jackson Laboratory 028555 mouse
DietGel 76A Clear H2O 70-01-5022 general supply
Dulbecco’s Modified Eagle’s Medium (DMEM) - high glucose Sigma Aldrich D6429 Medium
eBioscience 1X RBC Lysis Buffer Thermo fisher Scientific 00-4333-57 Solution
Falcon 100 mm TC-Treated Cell Culture Dish Life Sciences 353003 general supply
Falcon 5 mL round bottom polystyrene test tube Life Sciences 352054 general supply
Falcon 50 mL Conical Centrifuge Tubes Fisher Scientific 352098 general supply
Falcon 6 Well Clear Flat Bottom TC-Treated Multiwell Cell Culture Plate Life Science 353046 general supply
Fisherbrand microhematocrit capillary tubes Thermo Fisher Scientific 22-362566 general supply
Fisherbrand sterile cell strainers, 70 μm Fisher Scientific 22363548 general supply
Fixation Buffer BD Bioscience 554655 Solution
Guide-it Compete sgRNA Screening Systems Clontech 632636 Kit
Isothesia (Isoflurane) solution Henry Schein 29404 Solution
Lenti-X qRT-PCR Titration Kit Takara 631235 Kit
Lineage Cell Depletion Kit, mouse Miltenyi Biotec 130-090-858 Kit
Millex-HV Syringe Filter Unit, 0.45 mm Millipore Sigma SLHV004SL general supply
PBS pH7.4 (1X) Gibco 10010023 Solution
PEI MAX Polysciences 24765-1 Solution
Penicillin-Streptomycin Mixture Lonza 17-602F Solution
pLKO5.sgRNA.EFS.tRFP Addgene 57823 Plasmid
pMG2D Addgene 12259 Plasmid
Polybrene Infection/Transfection Reagent Sigma Aldrich TR-1003-G Solution
Polypropylene Centrifuge Tubes BECKMAN COULTER 326823 general supply
psPAX2 Addgene 12260 Plasmid
RadDisk – Rodent Irradiator Disk Braintree Scientific IRD-P M general supply
Recombinant Murine SCF Peprotech 250-03 Solution
Recombinant Murine TPO Peprotech 315-14 Solution
StemSpan SFEM STEMCELL Technologies 09600 Solution
TOPO TA cloning kit for sequencing with One Shot TOP10 Chemically Competent E. Thermo fisher Scientific K457501 Kit
Zombie Aqua Fixable Viability Kit BioLegend 423102 Solution
APC-anti-mouse Ly6C (Clone AL-21) BD Biosciences 560599 Antibodies
APC-Cy7-anti-mouse CD45R (RA3-6B2) BD Biosciences 552094 Antibodies
BV711-anti-mouse CD3e (Clone 145-2C11) Biolegend 100349 Antibodies
PE-Cy7-anti-mouse CD115 (Clone AFS98) eBioscience 25-1152-82 Antibodies
PerCP-Cy5.5-anti-mouse Ly6G (Clone 1A8) BD Biosciences 560602 Antibodies
FITC-anti-mouse CD4 (Clone RM4-5) Invitrogen 11-0042-85 Antibodies
BV510-anti-mouse CD8a (Clone 53-6.7) Biolegend 100752 Antibodies